Glycopegylation methods and proteins/peptides produced by the methods

Abstract

The invention includes methods and compositions for remodeling a peptide molecule, including the addition or deletion of one or more glycosyl groups to a peptide, and / or the addition of a modifying group to a peptide.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a Continuation of U.S. application Ser. No. 10 / 552,896 filed Jun. 8, 2006, now abandoned, which is a U.S. National Phase of PCT Application No. PCT / US2004 / 011494 filed Apr. 9, 2004, which application claims priority under 35 USC §120 to U.S. application Ser. No. 10 / 411,012 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,265,084, U.S. application Ser. No. 10 / 411,026 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,795,210, U.S. application Ser. No. 10 / 410,962 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,173,003, U.S. application Ser. No. 10 / 411,049 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,297,511, U.S. application Ser. No. 10 / 410,930 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,226,903, U.S. application Ser. No. 10 / 410,897 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,179,617, U.S. application Ser. No. 10 / 410,997 filed Apr. 9, 2003, issued as U.S. Pat. No. 7,157,277, U.S. application Ser. No. 10 / 411,044 filed Apr. 9, ...

AI Technical Summary

Benefits of technology

[0078]contacting the peptide with at least one glycosyltransferase and at least one glycosyl donor under conditions suitable to transfer at least one glycosyl donor to the truncated glycan, thereby remodeling the peptide.

[0084]contacting the peptide with at least one glycosyltransferase and at least one glycosyl donor under conditions suitable to transfer at least one glycosyl donor to the truncated glycan, thereby remodeling the peptide.

Problems solved by technology

The alteration of one or more of these characteristics greatly affects the efficacy of a peptide in its natural setting, and also affects the efficacy of the peptide as a therapeutic agent in situations where the peptide has been generated for that purpose.

Peptides whose glycans do not contain terminal sialic acid residues are rapidly removed from the circulation by the liver, an event which negates any potential therapeutic benefit of the peptide.

A major factor, which has limited the use of therapeutic peptides is the immunogenic nature of most peptides.

Other deficiencies of therapeutic peptides include suboptimal potency and rapid clearance rates.

The tetrasaccharide sialyl Lewis X was then enzymatically rebuilt on the remaining GlcNAc anchor site on the now homogenous protein by the sequential use of β-1,4-galactosyltransferase, α-2,3-sialyltransferase and α-1,3-fucosyltransferase V. However, while each enzymatically catalyzed step proceeded in excellent yield, such procedures have not been adapted for the generation of glycopeptides on an industrial scale.

Further, the methods of cell surface modification are not utilized for the enzymatic incorporation preformed modified glycosyl donor moiety into a peptide.

Moreover, none of the cell surface modification methods are practical for producing glycosyl-modified peptides on an industrial scale.

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